Clarke:Constructing vectors

1. Streak selective plate from glycerol stocks of E. coli that contain the vector plasmids and incubate overnight. Plates stored at 4 °C.
2. Pick colony from the plate and put in LB + antibiotic overnight.
3. Spin down cells, run miniprep kit to isolate plasmid DNA. Plasmids stored in buffer EB at -20 °C.
4. Take 1 μg of plasmid DNA, digest with restriction enzymes to give sticky ends that match the insert. If enzymes are heat inactivated, can store at -20 °C. (?)
5. Separate vector from cutout by gel electrophoresis. Gel slice stored at 4 °C.
6. Use gel extraction kit to recover vector. Vector stored in buffer EB at -20 °C.